1 Department of Ophthalmology, University of Florida, Gainesville, Florida.
2 Department of Ophthalmology, University of Alabama at Birmingham, Birmingham, Alabama.
Hum Gene Ther. 2019 May;30(5):571-589. doi: 10.1089/hum.2018.193. Epub 2018 Dec 20.
Mutations in , the gene encoding retinal guanylate cyclase-1 (retGC1), are the leading cause of autosomal dominant cone-rod dystrophy (CORD6). Significant progress toward clinical application of gene replacement therapy for Leber congenital amaurosis (LCA) due to recessive mutations in (LCA1) has been made, but a different approach is needed to treat CORD6 where gain of function mutations cause dysfunction and dystrophy. The CRISPR/Cas9 gene editing system efficiently disrupts genes at desired loci, enabling complete gene knockout or homology directed repair. Here, adeno-associated virus (AAV)-delivered CRISPR/Cas9 was used specifically to edit/disrupt this gene's early coding sequence in mouse and macaque photoreceptors , thereby knocking out retGC1 expression and demonstrably altering retinal function and structure. Neither preexisting nor induced Cas9-specific T-cell responses resulted in ocular inflammation in macaques, nor did it limit editing. The results show, for the first time, the ability to perform somatic gene editing in primates using AAV-CRISPR/Cas9 and demonstrate the viability this approach for treating inherited retinal diseases in general and CORD6 in particular.
突变 ，编码视网膜鸟苷酸环化酶-1（retGC1）的基因，是常染色体显性圆锥体-杆体营养不良（CORD6）的主要原因。由于 （LCA1）隐性突变导致莱伯先天性黑矇（LCA）的基因替代治疗已取得显著进展，但需要采用不同的方法来治疗 CORD6，因为功能获得性突变会导致功能障碍和营养不良。CRISPR/Cas9 基因编辑系统可在所需基因座高效地破坏基因，从而实现完全基因敲除或同源定向修复。在此，腺相关病毒（AAV）递送的 CRISPR/Cas9 被专门用于编辑/破坏该基因在小鼠和猕猴感光细胞中的早期编码序列，从而敲除 retGC1 的表达，并明显改变视网膜功能和结构。既没有预先存在的 Cas9 特异性 T 细胞反应，也没有诱导 Cas9 特异性 T 细胞反应导致灵长类动物的眼内炎症，也没有限制编辑。结果首次表明，使用 AAV-CRISPR/Cas9 在灵长类动物中进行体细胞基因编辑的能力，并证明了该方法治疗遗传性视网膜疾病，特别是 CORD6 的可行性。
